title: The generation of biomolecular patterns in highly porous collagen-GAG scaffolds using direct photolithography.
abstract: The extracellular matrix (ECM) is a complex organization of structural proteins found within tissues and organs. Heterogeneous tissues with spatially and temporally modulated properties play an important role in organism physiology. Here we present a benzophenone (BP) based direct, photolithographic approach to spatially pattern solution phase biomolecules within collagen-GAG (CG) scaffolds and demonstrate creation of a wide range of patterns composed of multiple biomolecular species in a manner independent from scaffold fabrication steps. We demonstrate the ability to immobilize biomolecules at surface densities of up to 1000 ligands per square micron on the scaffold strut surface and to depths limited by the penetration depth of the excitation source into the scaffold structure. Importantly, while BP photopatterning does further crosslink the CG scaffold, evidenced by increased mechanical properties and collagen crystallinity, it does not affect scaffold microstructural or compositional properties or negatively influence cell adhesion, viability, or proliferation. We show that covalently photoimmobilized fibronectin within a CG scaffold significantly increases the speed of MC3T3-E1 cell attachment relative to the bare CG scaffold or non-specifically adsorbed fibronectin, suggesting that this approach can be used to improve scaffold bioactivity. Our findings, on the whole, establish the use of direct, BP photolithography as a methodology for covalently incorporating activity-improving biochemical cues within 3D collagen biomaterial scaffolds with spatial control over biomolecular deposition.
authors.affiliation: Dept. of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
authors.forename: Teresa A
authors.initials: TA
authors.lastname: Martin
authors.forename: Steven R
authors.initials: SR
authors.lastname: Caliari
authors.forename: Paul D
authors.initials: PD
authors.lastname: Williford
authors.forename: Brendan A
authors.initials: BA
authors.lastname: Harley
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Bailey
mesh_terms: D000818:Animals; D002460:Cell Line; D003094:Collagen / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000737:chemistry*; D051379:Mice; D016062:Porosity; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D006025:Glycosaminoglycans; D003094:Collagen